Pancreatic cancer is the fourth leading cause of cancer-related deaths in US. Epidemiological data strongly suggest that the heavy drinking is a risk factor for pancreatic cancer. However, the molecular mechanism by which alcohol toxicity damages pancreas is not well understood. The main goal of the application is to examine the molecular mechanisms by which chronic alcohol consumption induces cellular transformation of human normal pancreatic ductal epithelial cells. Genetic variability and environmental exposures can modify the risk of pancreatic cancer. Cellular transformation is regulated by several genetic and epigenetic events. Progenitor cells / cancer stem cells play significant role in cellular transformation, and their emergence is responsible for cancer initiation and progression. We have found that during ethanol-induced cellular transformation, SATB2 gene (a transcription factor) is induced and its induction is directly correlated with the rate of transformation of pancreatic ductal epithelial cells. SATB2 is required for normal mammalian development; however, it is not expressed in normal adult cells. Inappropriate activation of SATB2 gene may cause malignant cellular transformation. SATB2 regulates transcription of pluripotency maintaining factors which are required for sustaining the stem cells capacity for self-renewal. Therefore, understanding the biology of ethanol-induced cellular transformation of normal pancreatic ductal epithelial cells is very significant for the management of pancreatitis and pancreatic cancer. We hypothesize that ethanol toxicity induces human pancreatic normal ductal epithelial cell transformation and enhances pancreatic carcinogenesis by up-regulating SATB2, and inhibition of SATB2 expression in ethanol-transformed cells suppresses their tumorigenic potential. Aim 1. To examine the molecular mechanisms by which ethanol induces cellular transformation of pancreatic normal ductal epithelial cells in vitro. Aim 2. To examine whether SATB2 mediates ethanol-induced cellular transformation. Aim 3. To assess whether induction or overexpression of SATB2 in human pancreatic normal ductal epithelial cells mediates pancreatic carcinogenesis in Balb C Nude mice. Aim 4. To examine whether chronic ethanol exposure regulates pancreatic carcinogenesis in vivo. The molecular mechanisms by which ethanol induces transformation of pancreatic normal ductal epithelial cells will be examined using qRT-PCR, ChIP-Seq, Western blot analysis, RNA-Seq and miRNA arrays. The expression of SATB2 in the pancreas isolated from human normal and alcoholic individuals will be compared. Gain and loss of SATB2 functional studies will be performed in vitro and in vivo. Ethanol, acetaldehyde, ADH, ALDH and CYP2E1 in the blood and tissues will be measured. Finally, KrasG12D/SATB2+/+, and KrasG12D/SATB2-/- mice will be generated, for the first time, to examine the involvement of SATB2 in ethanol-induced carcinogenesis. In conclusion, our novel studies will provide significant information about the ethanol-induced cellular transformation leading to pancreatic cancer.